Augmentation of cytokine production and mucosal immunity in gut-associated lymphoid tissue by the probiotics.

益生菌增强肠道相关淋巴组织中细胞因子的产生和粘膜免疫。

基本信息

批准号：
09660285
负责人：
KITAZAWA Haruki
金额：
$ 2.05万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

The Lactobacillus acidophilus group, a member of gastrointestinal lactic acid bacteria (LAB) widely used in dairy foods, is expected to develop as a probiotic LAB because of the ability to stimulate host immune responses. We have evaluated and assessed the mitogenic responses of the LAB to lymphocytes as an incidence of biological activity among them. The present study was conducted to determine the mitogenic factors from the LAB, particularly focusing on mitogenic DNAs in the LAB.In 12 of 16 strains of Lactobacillus acidophilus group LAB, the DNA components significantly stimulated mitogenic responses to murine splenocytes. The mitogenic activity to lymphocytes from peyer's patches, a gut-associated lymphoid tissue, was also observed in some DNAs. The active DNA (DNA1131) of Lactobacillus gasseri JCM1131^T was selected to analyze the active components. Plasmid DNAs from the positive clones obtained by suctioning, using the pUG119 vector ligated with 1131DNA partially digested with Sau … More 3Al, were purified by the precipitation method with polyethylene glycol. The plasmid DNA templates were amplified by using the polymerase chain reaction (POR) with primers for the multicloning site of the vector. In the 108 of the 321 positive colonies significant mitogenic activity was detected. The active nucleotides which stimulated mitogenic response were sequenced by a DNA sequencer MODEL 4000(Ll-COR, Lincoln, USA). Ten high homologous DNA motifs were found in the active DNA, but not in the non-active DNA.Two DNA motifs of chemically synthesized the DNA motifs induced a mitogenic response and was characterized as a B-cell specific mitogen. The binding of the active DNA clone and motifs to B lymphocytes were demonstrated by scatchard analysis and laser nicroscopy. The active DNA clone and motif augmented CD69/86 induction on B lymphocytes of spleen and peyer's patches. These also augmented induction of cytokine (IFNa. IFNy, TNRx) mRNA in splenocytes and peyer's patches. These findings indicated that a novel DNA motifs inducing lymphocyte activation triggering mitogenic response and , different from CpG motifs from E coli recently reported as a B-cell mitogen, exists in DNAs from L.Gasseri. Less

嗜酸乳杆菌是广泛用于乳制品中的胃肠道乳酸菌（LAB）的一员，由于能够刺激宿主免疫反应，因此有望发展为益生菌 LAB。我们已经评估和评估了 LAB 的促有丝分裂反应。本研究旨在确定 LAB 中的促有丝分裂因子，特别关注 LAB 中的促有丝分裂 DNA。在 16 个中的 12 个中。在嗜酸乳杆菌 LAB 菌株中，其 DNA 成分显着刺激了小鼠脾细胞的促有丝分裂反应，在一些加氏乳杆菌的 DNA 中也观察到了派伊尔氏淋巴结（一种肠道相关淋巴组织）的促有丝分裂活性。选择JCM1131^T，使用抽吸获得的阳性克隆来分析质粒DNA的活性成分。 pUG119 载体与用 Sau … More 3Al 部分消化的 1131DNA 连接，通过聚乙二醇沉淀法纯化质粒 DNA 模板，并使用聚合酶链式反应 (POR) 和载体多克隆位点的引物进行扩增。 321个阳性菌落中的108个被检测到显着的促有丝分裂活性。刺激促有丝分裂反应的活性核苷酸通过DNA测序仪进行了测序。 MODEL 4000（Ll-COR，Lincoln，USA）在活性DNA中发现了10个高度同源的DNA基序，但在非活性DNA中没有发现。化学合成的DNA基序中的两个DNA基序诱导有丝分裂反应，并被表征为通过斯卡查德分析和激光显微镜证实了活性 DNA 克隆和基序与 B 淋巴细胞的结合，活性 DNA 克隆和基序增强了 CD69/86 诱导。这些还增强了脾细胞和派尔氏集结中细胞因子（IFNa、IFNy、TNRx）mRNA 的诱导。这些发现表明，诱导淋巴细胞活化的新 DNA 基序与 CpG 基序不同。最近报道，大肠杆菌是一种 B 细胞有丝分裂原，存在于 L.Gasseri Less 的 DNA 中。